A fluorescence polarization assay using oligonucleotide probes for the rapid detection of verotoxin-producing Escherichia coli

被引：20
作者：
Ohiso, I
Tsuruoka, M
Iida, T
Honda, T
Karube, I
机构：
[1] Nishikawa Rubber Co Ltd, Diversified Prod Div, Nishi Ku, Hiroshima 7338510, Japan
[2] Adv Sci & Technol Lab, Asaminami Ku, Hiroshima 7313162, Japan
[3] Osaka Univ, Res Inst Microbial Dis, Dept Bacterial Infect, Suita, Osaka 5650871, Japan
[4] Univ Tokyo, Adv Sci & Technol Res Ctr, Meguro Ku, Tokyo 1538904, Japan
来源：
JOURNAL OF BIOTECHNOLOGY | 2000年 / 81卷 / 01期
关键词：
fluorescence polarization; verotoxin; Escherichia coli; verotoxin-producing E. coli; VT2; DNA probe;
D O I：
10.1016/S0168-1656(00)00261-3
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
A hybridization assay using fluorescence polarization was combined with the asymmetric polymerase chain reaction (PCR) in a method for the detection of the verotoxin type 2 gene of verotoxin-producing Escherichia coli. Six oligonucleotide probes labeled with FITC were designed and evaluated. One of these gave a detection limit of 10(3) colony forming units per assay, and assay results could be obtained within 5 min after PCR. It appears that the detection limit was restricted mainly by the extent and fidelity of PCR amplification, rather than by the sensitivity of the fluorescence polarization technique, indicating that good probe design facilitates the rapid detection of the PCR product. The fluorescence polarization assay, in conjunction with DNA amplification by PCR, is a powerful and widely applicable method for the rapid and sensitive detection of oligonucleotide sequences. (C) 2000 Elsevier Science B.V. All rights reserved.
引用
收藏
页码：15 / 25
页数：11
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